Tutorials

Obtaining Reliable PCR from Seed DNA

DNA from Monocot Seeds

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Figure 1. PCR of DNA extracted from monocot seeds

QuickExtract Seed Solution was used to screen monocot seeds, which often yield DNA that gives inconsistent amplification results by standard methods. Approximately 10 mg of a variety of ground or fragmented monocot crop seeds were processed using 100 µL of the solution for each sample. Aliquots of the undiluted extracts were amplified by PCR with conserved intron-spanning primers. The expected amplicon was produced in all cases (Figure 1).

Next, we used approximately 10 mg of corn flour in each conical well of a 96-well plate and extracted DNA with 100 µL of QuickExtract Seed Solution as before. The plate was centrifuged for five minutes at 3,000 rpm. A 0.5-µL aliquot of each extract was added to the PCR mixture in a separate 96-well plate containing the PlantAmp PCR PreMix and primers for a universal monocot gene. The amplified locus, BRSC3, spans an intron and is highly conserved among monocot plants. The expected band of around 250 bp was amplified in all 96 samples; representative examples are shown in Figure 2.

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